Zinc Finger Protein-Protein Interactions

锌指蛋白-蛋白质相互作用

• 批准号: 7026436
• 负责人: J. KEITH JOUNG
• 金额: $ 30.76万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7026436
• 关键词: SDS polyacrylamide gel electrophoresis; X ray crystallography; biophysics; gene expression; genetic regulation; protein protein interaction; protein structure; protein structure function; transcription factor

DESCRIPTION (provided by applicant): Protein-protein interactions among transcription factors play critical roles in the regulation of gene expression in normal and disease states. This application proposes to study protein-protein interactions mediated by Cys2His2 zinc finger proteins, the most abundant class of transcription factors in humans. In particular, we are focusing our studies on dimerization zinc finger (DZF) domains found in members of the Ikaros family of transcription factors and related homologues. DZF domains provide a model system for studying Cys2His2 zinc finger-mediated protein-protein interactions. The long-term goals of this proposal are: to gain a detailed biophysical understanding of how Cys2His2 zinc fingers mediate specific interactions with a wide variety of different proteins; to utilize this knowledge to devise strategies to enhance, disrupt, or predict such interactions; and to engineer synthetic zinc fingers with novel interaction specificities that could serve as useful modules for creating artificial regulatory circuits with potential applications in biotechnology and therapeutics. The specific aims of this application are: 1) to define and model the Ikaros, Pegasus, and Hunchback DZF domain interfaces using mutational analysis, genetic selection methods, and molecular graphics tools; 2) to determine high-resolution structures of dimeric Ikaros, Pegasus, and Hunchback DZF domain protein-protein complexes using X-ray crystallography; 3) to use genetic selection to identify synthetic, obligate heterodimeric DZF domains and to test the use of DZF domains as "interaction modules" for assembling heterologous protein complexes in eukaryotic cells.

描述（由申请人提供）：转录因子之间的蛋白质 - 蛋白质相互作用在正常和疾病状态中基因表达的调节中起关键作用。该应用建议研究由CYS2HIS2锌指蛋白介导的蛋白质 - 蛋白质相互作用，这是人类中最丰富的转录因子类别。特别是，我们将研究重点放在Ikaros转录因子和相关同源物的Ikaros家族成员中发现的二聚化锌指（DZF）领域。 DZF结构域提供了一个模型系统，用于研究Cys2HIS2锌指介导的蛋白质 - 蛋白质相互作用。该提案的长期目标是：对Cys2HIS2锌指的方式获得详细的生物物理理解，以介导特定的相互作用与多种不同蛋白质；利用这些知识来制定策略来增强，破坏或预测此类相互作用；并使用新型相互作用特异性来设计合成锌指的手指，这些特异性可以用作创建具有生物技术和治疗剂潜在应用的人工调节回路的有用模块。该应用程序的具体目的是：1）使用突变分析，遗传选择方法和分子图形工具来定义和建模Ikaros，Pegasus和Hunchback DZF域界面； 2）使用X射线晶体学确定二聚体Ikaros，Pegasus和Hunchback DZF域蛋白质蛋白复合物的高分辨率结构； 3）使用遗传选择来识别合成的，强性的异二聚体DZF结构域，并测试DZF结构域作为“相互作用模块”用于组装真核细胞中异源蛋白复合物的“相互作用模块”。